Component analysis method and component analysis device

ABSTRACT

The present disclosure provides component analysis methods including a measurement process and an analysis process.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority under 35 USC 119 from Japanese PatentApplication No. 2018-155960, filed on Aug. 23, 2018, the disclosure ofwhich is incorporated by reference herein.

BACKGROUND Technical Field

The present disclosure relates to a component analysis method and acomponent analysis device that use continuous sample introduction.

Related Art

In a component analysis system using continuous sample introduction suchas capillary electrophoresis, there is technology in which a curve as abase waveform is plotted with detection data such as optical absorbanceacquired by a detector along a vertical axis and time along thehorizontal axis, and component analysis is performed using adifferentiated waveform such as an electropherogram that is obtained bydifferentiating the base waveform with respect to time.

Each peak appearing in the differentiated waveform corresponds to acomponent contained in the introduced sample. Moreover, each componentcan be identified by a time difference when a top of each peak appears.Furthermore, an area occupied by each of the peaks in the differentiatedwaveform is employed as an indicator of an amount of each correspondingcomponent in the sample. For example, a differentiated waveform in ahemoglobin assay system obtained by continuous sample introduction witha blood sample shows a shape such as that illustrated in Japanese PatentApplication Laid-Open (JP-A) No. 2018-72336.

In a component analysis system using continuous sample introduction suchas in capillary electrophoresis, components are identified by the peaksappearing in the differentiated waveform obtained by differentiating theoptical absorbance curve along the time axis as described above, and thecomponents are relatively quantified by the area occupied by theportions at these peaks in the differentiated waveform. In such asystem, a valley portion (hereafter referred to as “bottom”) givenbetween the peaks is often used as the boundary between the peaks.However, such a bottom is frequently indistinct. In particular, when twopeaks are merged, a lower peak is sometimes absorbed in a higher peak,it is difficult to distinguish between the two peaks and, in such acase, it is difficult or impossible to identify the bottom.

SUMMARY

An exemplary embodiment of the present disclosure is a componentanalysis method including a measurement process in which a samplesolution that is continuously introduced into a flow path is separatedinto a plurality of components inside the flow path and is opticallymeasured over time at a measurement position on the flow path to obtainoptical measurement values, and an analysis process in which theplurality of components contained in a sample is analyzed based on theoptical measurement values. The analysis process includes a basewaveform acquisition process in which a base waveform is acquired byplotting the optical measurement values along a time axis on atwo-dimensional plane, a measurement value differentiation process inwhich a measurement value differentiated waveform is acquired, which isa waveform obtained by differentiating the base waveform along an axisof the optical measurement values orthogonal to the time axis, and ameasurement value boundary determination process in which the opticalmeasurement values corresponding to peaks in the measurement valuedifferentiated waveform are determined to be separation boundariesbetween adjacent components of the plural components.

The exemplary embodiment of the present disclosure enables, in acomponent analysis system using continuous sample introduction, aboundary to be definitively determined even in cases in which it isdifficult to determine a distinct boundary between two peaks in adifferentiated waveform.

BRIEF DESCRIPTION OF THE DRAWINGS

Exemplary embodiments will be described in detail based on the followingfigures, wherein:

FIG. 1 is a system schematic diagram that illustrates a componentanalysis system that can be used to execute a component analysis methodof an exemplary embodiment;

FIG. 2 is a plan view that illustrates an analysis chip used in theanalysis system of FIG. 1;

FIG. 3 is a cross-section that is taken along line in FIG. 2;

FIG. 4 is a block diagram that illustrates a hardware configuration of acontrol section;

FIG. 5 is a block diagram that represents a functional configuration ofa component analysis device;

FIG. 6 is a block diagram that represents a functional configuration ofan analysis unit of a first aspect;

FIG. 7 is a flowchart that schematically illustrates a componentanalysis method of the first aspect;

FIG. 8 illustrates an example of a base waveform as a solid line;

FIG. 9 illustrates a time differentiated waveform with respect to thebase waveform of FIG. 8 as a dashed line;

FIG. 10 illustrates an appended dotted line of a measurement valuedifferentiated waveform with respect to the base waveform of FIG. 9;

FIG. 11 is a block diagram that illustrates a functional configurationof an analysis unit of a second aspect;

FIG. 12 is a flowchart that schematically illustrates a componentanalysis method of the second aspect;

FIG. 13 is a block diagram that illustrates a functional configurationof an analysis unit of a third aspect;

FIG. 14 is a flowchart that schematically illustrates a componentanalysis method of the third aspect;

FIG. 15 illustrates a measurement value differentiated waveform withrespect to the base waveform of FIG. 8 illustrated as a dotted line;

FIG. 16 is a block diagram that illustrates a functional configurationof an analysis unit of a fourth aspect;

FIG. 17 is a flowchart that schematically illustrates a componentanalysis method of the fourth aspect;

FIG. 18 is chart appended with a reciprocal differentiated waveform withrespect to the time differentiated waveform of FIG. 9;

FIG. 19 is a flowchart that represents a functional configuration of ananalysis unit of a fifth aspect;

FIG. 20 is a flowchart that schematically represents a componentanalysis method of the fifth aspect;

FIG. 21A to FIG. 21D are charts that illustrate effects of high and lowambient temperatures and high and low concentrations of a specimenduring measurement on a time differentiated waveform obtained by a timedifferentiation process: FIG. 21A illustrates a case of a lowconcentration sample at the ambient temperature of 23° C., FIG. 21Billustrates a case of a high concentration sample at the ambienttemperature of 23° C., FIG. 21C illustrates a case of a lowconcentration sample at the ambient temperature of 8° C., and FIG. 21Dillustrates a case of a high concentration sample at the ambienttemperature of 8° C.; and

FIG. 22A to FIG. 22D are charts that illustrate effects of high and lowambient temperatures and high and low concentrations of a specimenduring measurement on a time differentiated waveform obtained by a timedifferentiation process: FIG. 22A illustrates a case of a lowconcentration sample at the ambient temperature of 23° C., FIG. 22Billustrates a case of a high concentration sample at the ambienttemperature of 23° C., FIG. 22C illustrates a case of a lowconcentration sample at the ambient temperature of 8° C., and FIG. 22Dillustrates a case of a high concentration sample at the ambienttemperature of 8° C.

DETAILED DESCRIPTION

Explanation follows regarding exemplary embodiments of the presentdisclosure, with reference to the drawings as appropriate.

Component Analysis System

FIG. 1 illustrates a schematic configuration of a component analysissystem A1 in which a component analysis method according to the presentexemplary embodiment can be executed. The component analysis system A1is configured including a component analysis device 1 and an analysischip 2. The component analysis system A1 is a system for executing ananalysis method using capillary electrophoresis with a sample Sa as atarget. The sample Sa may include any component that is able to beanalyzed by capillary electrophoresis, and a property of the sample Samay be liquid, solid, or gas as long as the sample Sa is soluble to amedium. Explanation follows regarding an example in the presentexemplary embodiment in which the sample Sa is blood collected from ahuman body.

Examples of components to be analyzed among the components contained inthe sample Sa include hemoglobin (Hb), albumin (Alb), globulin (α1, α2,β, and γ-globulin), fibrinogen, or the like. Examples of the hemoglobinreferred to above include several types of hemoglobin, such as normalhemoglobin (HbA), hemoglobin variants (HbA1c, HbC, HbD, HbE, HbS, etc.),fetal hemoglobin (HbF), and the like. These components are preferablyanalyzed using capillary electrophoresis since mutations in the aminoacids as configuration elements thereof are readily reflected in proteinmolecules electrically. In the component analysis system A1 according tothe present exemplary embodiment, separation boundaries between aplurality of components are determined, which are a plurality ofhemoglobin variants as a plurality of mutants of the same kinds ofproteins.

Depending on the components subjected to analysis, the sample may bepre-processed with suitable reagents, or may be subjected to preliminaryseparation processing using a different method (such as, for example, achromatography method).

The analysis chip 2 holds the sample Sa, and provides a place forperforming analysis on the target sample Sa in a state in which theanalysis chip 2 has been loaded in the component analysis device 1. Inthe present exemplary embodiment, the analysis chip 2 is configured by aso-called disposable type of analysis chip, which is intended to bedisposed of after a single analysis has been completed. As illustratedin FIG. 2 and FIG. 3, the analysis chip 2 includes a body 21, a mixingreservoir 22, an introduction reservoir 23, a filter 24, a dischargereservoir 25, an electrode reservoir 26, a flow path 27, and acommunication flow path 28. FIG. 2 is a plan view of the analysis chip2, and FIG. 3 is a cross-section taken along III-III in FIG. 2. Notethat the analysis chip 2 is not limited to the disposable type, and maybe a chip that is used for several analyses. Moreover, the componentanalysis system according to the present exemplary embodiment is notlimited to a configuration in which the analysis chip 2 is provided as aseparate body to be loaded into the component analysis device 1. Thecomponent analysis system according to the present invention may beconfigured with a functional part that accomplishes similarfunctionality to that of the analysis chip 2 and is built into thecomponent analysis device 1.

The body 21 is a base for the analysis chip 2. The material of the body21 is not particularly limited and examples thereof include glass, fusedsilica, plastic, and the like. In the present exemplary embodiment, thebody 21 is formed from separate bodies, these being an upper portion 2Aand a lower portion 2B illustrated in FIG. 3, in a configuration inwhich the upper portion 2A and the lower portion 2B are joined to eachother. Note that there is no limitation thereto, and for example, thebody 21 may be formed as a single integrated unit.

The mixing reservoir 22 is a site where the sample Sa and a diluent Ldare mixed. The mixing reservoir 22 is, for example, configured as arecess open to the upper side by a through hole formed in the upperportion 2A of the body 21. The introduction reservoir 23 is a reservoirinto which a sample solution is introduced which is obtained from mixingthe sample Sa and the diluent Ld together in the mixing reservoir 22.The introduction reservoir 23 is, for example, configured as a recessopen to the upper side by a through hole formed in the upper portion 2Aof the body 21.

The filter 24 is provided to an opening of the introduction reservoir23, with the opening serving as an example of an introduction path tothe introduction reservoir 23. The specific configuration of the filter24 is not limited, and a preferable example thereof is a celluloseacetate membrane filter (manufactured by ADVANTEC and having a 0.45 μmpore diameter).

The discharge reservoir 25 is a reservoir that is positioned on thedownstream side of the flow path 27. The discharge reservoir 25 is, forexample, configured as a recess open to the upper side by a through holeformed in the upper portion 2A of the body 21. The electrode reservoir26 is a reservoir into which an electrode 31 is inserted in capillaryelectrophoresis. The electrode reservoir 26 is, for example, configuredas a recess open to the upper side by a through hole formed in the upperportion 2A of the body 21. The communication flow path 28 connects theintroduction reservoir 23 and the electrode reservoir 26 together, andconfigures a contiguous path between the introduction reservoir 23 andthe electrode reservoir 26.

The flow path 27 is formed as a capillary tube connecting theintroduction reservoir 23 and the discharge reservoir 25 together, andis a place where electroosmotic flow (EOF) occurs during capillaryelectrophoresis. The flow path 27 is, for example, configured as agroove formed in the lower portion 2B of the body 21. Note that recessesor the like may be formed in the body 21 as appropriate to promoteirradiation of light onto the flow path 27 and to promote output oflight that has passed through the flow path 27. The size of the flowpath 27 is not particularly limited, and as an example thereof, the flowpath 27 has a width of 25 μm to 100 μm, a depth of 25 μm to 100 μm, anda length of 5 mm to 150 mm. The overall size of the analysis chip 2 isset as appropriate according to the size of the flow path 27 and thesize, placement, and so on, of the mixing reservoir 22, the introductionreservoir 23, the discharge reservoir 25, and the electrode reservoir26.

Note that the analysis chip 2 configured as described above is merely anexample, and any analysis chip with a configuration that enablescomponent analysis to be performed by capillary electrophoresis may beappropriately employed therefor.

The component analysis device 1 performs analysis processing on thesample Sa, in a state in which the analysis chip 2 spotted with thesample Sa is loaded in the component analysis device 1. The componentanalysis device 1 includes an electrode 31, an electrode 32, a lightsource 41, an optical filter 42, a lens 43, a slit 44, a detector 45, adispenser 6, a pump 61, a diluent reservoir 71, a migration liquidreservoir 72, and a control section 8. Note that the light source 41,the optical filter 42, the lens 43, and the detector 45 configure ameasurement section 40 in the present exemplary embodiment.

The electrode 31 and the electrode 32 are for applying a predeterminedvoltage to the flow path 27 during capillary electrophoresis. Theelectrode 31 is inserted into the electrode reservoir 26 of the analysischip 2, and the electrode 32 is inserted into the discharge reservoir 25of the analysis chip 2. The voltage applied to the electrode 31 and theelectrode 32 is not particularly limited, and may, for example, be from0.5 kV to 20 kV.

The light source 41 is a part that emit light to measure opticalabsorbance as an optical measurement value during capillaryelectrophoresis. The light source 41 is provided, for example, with anLED chip that emits light of a predetermined wavelength range. Theoptical filter 42 attenuates light of predetermined wavelengths in thelight from the light source 41 and transmits the light of otherwavelengths therein. The lens 43 is focuses light that is transmittedthrough the optical filter 42 onto an analysis site of the flow path 27of the analysis chip 2. The slit 44 excludes excess light from the lightfocused using the lens 43, which might otherwise cause scattering andthe like.

The detector 45 receives light from the light source 41 that istransmitted through the flow path 27 of the analysis chip 2, and isconfigured including a photodiode, a photo IC, or the like.

In this manner, a light path is a route through which the light emittedby the light source 41 to the detector 45. An optical measurement valuefor the solution (namely, either a sample solution, a migration liquid,or a mixture solution thereof) flowing through the flow path 27 ismeasured at a measurement position 27A, which is the position where thislight path intersects with the flow path 27. Namely, the opticalmeasurement value of the sample solution is measured by the measurementsection 40 at the measurement position 27A on the flow path 27. Theoptical measurement value may, for example, be optical absorbance.Optical absorbance represents a degree to which light in the light pathis absorbed by the solution flowing through the flow path 27, and isexpressed as an absolute value of a value for the common logarithm of aratio of incident light intensity to transmitted light intensity. Insuch cases, a generic spectrophotometer may be employed as the detector45. Note that any optical measurement value, such as the value of thesimple transmitted light intensity, may be used in the present inventioninstead of the optical absorbance. The following explanation describesan example in which optical absorbance is used as the opticalmeasurement value.

The dispenser 6 dispenses desired amounts of the diluent Ld, a migrationliquid Lm, and the sample solution, and the dispenser 6 includes anozzle, for example. The dispenser 6 can be freely moved between aplurality of predetermined positions in the component analysis device 1using a drive mechanism, not illustrated in the drawings. The pump 61 isa draw source to the dispenser 6 and a purge source from the dispenser6. Moreover, the pump 61 may be used as a draw source and a purge sourcefor ports, not illustrated in the drawings, provided to the componentanalysis device 1. Such ports may be employed to fill the migrationliquid Lm or the like. Dedicated pumps may also be provided therefor,separate to the pump 61.

The diluent reservoir 71 is a reservoir for storing the diluent Ld. Thediluent reservoir 71 may be a reservoir that is permanently installed tothe component analysis device 1, or may be a container filled with apredetermined amount of the diluent Ld and loaded into the componentanalysis device 1. The migration liquid reservoir 72 is a reservoir forstoring the migration liquid Lm. The migration liquid reservoir 72 maybe a reservoir that is permanently installed to the component analysisdevice 1, or may be a container filled with a predetermined amount ofthe migration liquid Lm and loaded into the component analysis device 1.

The diluent Ld is mixed with the sample Sa to produce the samplesolution. The main agent of the diluent Ld is not particularly limited,and examples thereof include water and physiological saline. Apreferable example of the diluent Ld is a liquid including componentssimilar to those of the migration liquid Lm, described later. Moreover,additives may also be added as necessary to the diluent Ld in additionto the main agent.

The migration liquid Lm is filled into the discharge reservoir 25 andthe flow path 27 in an analysis process S20 employing electrophoresis,and is a medium to generate electroosmotic flow during capillaryelectrophoresis. Although the migration liquid Lm is not particularlylimited, an acid is preferably used therefor. The acid is, for example,citric acid, maleic acid, tartaric acid, succinic acid, fumaric acid,phthalic acid, malonic acid, or malic acid. The migration liquid Lmpreferably also includes a weak base. The weak base is, for example,arginine, lysine, histidine, tris, or the like. The pH of the migrationliquid Lm is, for example, in a range of pH 4.5 to pH 6. A type of abuffer in the migration liquid Lm is MES, ADA, ACES, BES, MOPS, TES,HEPES, or the like. Similar to the above explanation regarding thediluent Ld, additives may also be added to the migration liquid Lm asnecessary.

The control section 8 controls each section in the component analysisdevice 1. The control section 8 includes, as illustrated in a hardwareconfiguration in FIG. 4, a central processing unit (CPU) 81, read onlymemory (ROM) 82, random access memory (RAM) 83, and storage 84. Theseparts of the configuration are connected together so as to be capable ofcommunicating with each other through a bus 89.

The CPU 81 is a central processing unit that executes various programsto control each section. Namely, the CPU 81 reads a program from the ROM82 or the storage 84, and executes the program using the RAM 83 as aworking area. The CPU 81 performs control of the configuration shownabove and various computation processing according to the programrecorded in the ROM 82 or the storage 84.

The ROM 82 stores various programs and various data. The RAM 83 servesas a working area to temporarily store programs or data. The storage 84is configured by a hard disk drive (HDD), solid state drive (SSD), orflash memory, and is stored with various programs including an operatingsystem, and various data. In the present exemplary embodiment, a programto execute the component analysis method according to the presentexemplary embodiment and various data is stored in the ROM 82 or thestorage 84.

In order to execute the component analysis method according to thepresent exemplary embodiment, the component analysis device 1 executesvarious functions such as those illustrated in FIG. 5 using the hardwareresources and configuration parts described above. These functionsinclude, as well as the function of the measurement section 40, thefunction of an analysis unit 800 to analyze a plurality of componentscontained in the sample Sa based on the optical measurement valuesobtained by measuring the sample solution optically over time with themeasurement section 40. These functions are described later.

First Aspect

A component analysis method of the first aspect of the presentdisclosure includes: a measurement process S10 and an analysis processS20. In the measurement process S10, a sample solution that iscontinuously introduced into a flow path 27 is separated into aplurality of components inside the flow path 27 and is opticallymeasured over time at a measurement position 27A on the flow path 27 toobtain optical measurement values. In the analysis process S20, theplurality of components contained in a sample Sa is analyzed based onthe optical measurement values. The analysis process S20 includes: abase waveform acquisition process S21 in which a base waveform isacquired by plotting the optical measurement values along a time axis ona two-dimensional plane; a measurement value differentiation process S23in which a measurement value differentiated waveform is acquired, whichis a waveform obtained by differentiating the base waveform along anaxis of the optical measurement values orthogonal to the time axis; anda measurement value boundary determination process S24 in which theoptical measurement values corresponding to peaks in the measurementvalue differentiated waveform are determined to be separation boundariesbetween adjacent components of the plural components.

A component analysis device 1 of the present aspect includes: a flowpath 27 into which a sample solution is continuously introduced; ameasurement section 40 that optically measures the sample solution overtime, which is separated into a plurality of components inside the flowpath 27, at a measurement position 27A on the flow path 27 to obtainoptical measurement values; and an analysis unit 800 that analyzes theplurality of components contained in a sample Sa based on the opticalmeasurement values. The analysis unit 800 includes: a base waveformacquisition section 801 that acquires a base waveform by plotting theoptical measurement values along a time axis on a two-dimensional plane;a measurement value differentiation section 803 that acquires ameasurement value differentiated waveform that is a waveform obtained bydifferentiating the base waveform along an axis of the opticalmeasurement values orthogonal to the time axis; and a measurement valueboundary determination section 804 that determines optical measurementvalues corresponding to peaks in the measurement value differentiatedwaveform to be separation boundaries between adjacent components intothe plural components.

A functional configuration of the analysis unit 800 of the presentaspect is illustrated in FIG. 6. Explanation follows regarding thecomponent analysis method of the present aspect, with reference to aflowchart of FIG. 7.

In a measurement process S10 illustrated in FIG. 7, the sample solutionis continuously introduced into the flow path 27, the sample solution isseparated into the plurality of components in the flow path 27 byapplication of a voltage, and an optical measurement value is obtainedfor when the sample solution arrives at the measurement position 27Aprovided on a way along the flow path 27 by the measurement section 40.Specifically, the electrodes 31, 32 are respectively mounted at anupstream side and a downstream sides of the flow path 27 and the samplesolution is applied with the voltage therebetween, and is therebysubjected to electrophoresis in so-called capillary electrophoresis. Inother words, the optical measurement values are obtained using capillaryelectrophoresis in which a voltage is applied to the sample solutionthat is continuously introduced into a capillary tube as the flow path27 so as to separate the sample solution into the plurality ofcomponents.

For example, when a target component for analysis is hemoglobin asdescribed above, then placing a negative electrode as the electrode 31at the upstream side of the flow path 27 and placing a positiveelectrode as the electrode 32 at the downstream side of the flow path 27results in the analysis target component being migrated toward theelectrode 32 as the positive electrode due to surfaces of molecules ofthe hemoglobin being negatively charged. When this occurs, an electricalmigration speed varies according to a charged state of the moleculesurface. Namely, the stronger a negative charge on the molecule surfaceis, the faster the migration speed is. Thus, when the sample solution isintroduced into the flow path 27, the component of a higher migrationspeed earlier arrives at the measurement position 27A.

Then, at a time when the component of a lower migration speed arrives atthe measurement position 27A, the component of a higher migration speed,which is derived from the sample solution introduced later, also arrivesat the same time at the measurement position 27A. Namely, once thecomponent of a higher migration speed arrives at the measurementposition 27A in the flow path 27, the component continues to arrive atthe measurement position 27A as long as the sample solution iscontinuously introduced into the flow path 27. Moreover, the componentof a lower migration speed arrives at the measurement position 27Alater. However, once the component of a lower migration speed arrives atthe measurement position 27A, the component also continues to arrive atthe measurement position 27A as long as the sample solution iscontinuously introduced into the flow path 27.

In other words, the component of a higher migration speed arrives at themeasurement position 27A earlier, and thereafter each the component of alower migration speed arrives at the measurement position 27Acumulatively. Therefore, the optical absorbance measured as the opticalmeasurement value of the sample solution by the measurement section 40at the measurement position 27A chronologically shows a cumulative andmonotonous increase.

In a base waveform acquisition process S21 in the analysis process S20illustrated in FIG. 7, a base waveform acquisition section 801 in theanalysis unit 800 illustrated in FIG. 6 acquires a base waveform byplotting the optical absorbance values as the optical measurement valuesalong one axis (the Y axis, for example) of a two-dimensional planeagainst a time axis as another axis (the X axis, for example). The basewaveform is, for example, represented as a solid line on a graph such asillustrated in FIG. 8. The optical measurement values in the presentaspect may be acquired as data that are capable of being plotted on atwo-dimensional plane, and a graph based on such data does not actuallyneed to be drawn. This also similarly applies to other aspects mentionedbelow. Note that the base waveform may be referred to as a function inwhich the optical measurement value is plotted with time as a variable.

Viewing this base waveform along the time axis, the portions of highgradient indicated by solid line arrows in FIG. 8 are caused by anincrease in the optical measurement value due to a given componentarriving at the measurement position 27A for the first time. Moreover,the portions of low gradient indicated by dashed line arrows thereinindicate that the next component does not yet arrive at the measurementposition 27A. Namely, the portions of high gradient in the base waveformindicate the arrival of components in the sample solution at themeasurement position 27A. Such portions of high gradient in the basewaveform, as illustrated in a dashed line on a graph in FIG. 9, may berepresented by peak waveforms appearing in a waveform obtained bydifferentiating the base waveform along the time axis (hereinafterreferred to as a “time differentiated waveform”). Such peak waveformsmay be identified as being caused by each component. Note that on thetime axis, a component corresponding to a peak waveform that locatesmore to the left represents a component of a higher migration speed thanthat of a component corresponding to a peak waveforms that locates moreto the right.

When there are a plurality of peak waveforms in the time differentiatedwaveform, adjacent two peak waveforms are demarcated by a bottom B,which is a valley portion between the two peak waveforms. The peaks T ofthe peak waveforms are maximal values in the time differentiatedwaveform, and correspond to locations in the base waveform where slopesthereof show maximal values. On the other hand, the bottoms B areminimal values in the time differentiated waveform, and correspondingslopes in the base waveform show minimal values. In other words, eachportion of a steep slope in the base waveform corresponds to a peak T,and each portion of a gentle slope therein corresponds to a bottom B.

On the other hand, viewing the base waveform illustrated in FIG. 8 alongthe optical measurement value axis (Y axis), each portion correspondingto a peak T has a gentle slope, and each portion corresponding to abottom B has a steep slope.

A measurement value differentiation process S23 in the analysis processS20 illustrated in FIG. 7 focuses on this point. Namely, in themeasurement value differentiation process S23, a measurement valuedifferentiation section 803 in the analysis unit 800 illustrated in FIG.6 differentiates the base waveform along the optical measurement valueaxis, and acquires a measurement value differentiated waveform asillustrated by the dotted line in a graph in FIG. 10.

As illustrated in FIG. 10, the peaks T1′ to T6′ on the measurement valuedifferentiated waveform respectively correspond to the bottoms B1 to B6on the time differentiated waveform. Namely, in the measurement valuedifferentiated waveform, peaks and bottoms are taken over with respectto peaks and bottoms in the time differentiated waveform. In otherwords, the bottoms B1 to B6 in the time differentiated waveformrespectively appear clearly as the peaks T1′ to T6′ in the measurementvalue differentiated waveform. Note that the measurement valuedifferentiated waveform may be referred to as a function in which thetime of the base waveform is differentiated by the optical measurementvalue as a variable.

Then, in a measurement value boundary determination process S24 in theanalysis process S20 illustrated in FIG. 7, a measurement value boundarydetermination section 804 in the analysis unit 800 illustrated in FIG. 6determines optical measurement values S1 to S6 respectivelycorresponding to the peaks T1′ to T6′ to be separation boundariesbetween components.

The configuration of the first aspect as described above enables thebottoms B1 to B6, which are hitherto needed to be discriminated as thetwo ends of a peak waveform in a time differentiated waveform, can berespectively identified as the peaks T1′ to T6′ in the measurement valuedifferentiated waveform. Then, it is possible for the opticalmeasurement values S1 to S6 corresponding to the peaks T1′ to T6′ to berespectively determined to be the separation boundaries between thecomponents contained in the sample Sa.

Note that it is not necessary for acquiring the peaks T1′ to T6′ in themeasurement value differentiated waveform to refer to a timedifferentiated waveform at all. The reason why the time differentiatedwaveform is referenced in the above explanation and in FIG. 9 and FIG.10 is merely to explain the meaning of the peaks T1′ to T6′ in themeasurement value differentiated waveform.

Second Aspect

A component analysis method of a second aspect of the present disclosureis an augmented configuration of the component analysis method of thefirst aspect, wherein the analysis process S20 further includes: a timedifferentiation process S22 in which a time differentiated waveform isacquired, which is a waveform obtained by differentiating the basewaveform along the time axis; and an integral quantification process S28in which a value obtained by integrating the time differentiatedwaveform for an integration interval whose both ends are adjacent twointegration boundaries, wherein time points corresponding to theseparation boundaries are determined to be integration boundaries forthe time differentiated waveform, is calculated as a relative amount ofeach component in the sample Sa corresponding to the integrationinterval.

The component analysis device 1 in the present aspect is an augmentedconfiguration of the component analysis device 1 of the first aspect,wherein the analysis unit 800 further includes: a time differentiationsection 802 that acquires a time differentiated waveform that is awaveform obtained by differentiating the base waveform along the timeaxis; and an integral quantification section 808 that calculates, avalue obtained by integrating the time differentiated waveform for eachintegration interval whose both end are adjacent two integrationboundaries, wherein time points corresponding to the separationboundaries are determined to be integration boundaries for the timedifferentiated waveform, as a relative amount of each component in thesample Sa corresponding to the integration interval.

A functional configuration of the analysis unit 800 of the presentaspect is illustrated in FIG. 11. Explanation follows regarding thecomponent analysis method of the present aspect, with reference to aflowchart of FIG. 12.

The measurement process S10 illustrated in FIG. 12 is similar to that inthe first aspect.

In a base waveform acquisition process S21 of an analysis process S20illustrated in FIG. 12, a base waveform acquisition section 801 of theanalysis unit 800 illustrated in FIG. 11 acquires a base waveform byplotting the optical absorbance values as the optical measurement valuesalong one axis (the Y axis, for example) of a two-dimensional planeagainst a time axis as another axis (the X axis, for example). The basewaveform is, for example, represented as a solid line on a graph such asillustrated in FIG. 8. The base waveform acquisition process S21 issimilar to that of the first aspect.

Then, in a time differentiation process S22 in the analysis process S20illustrated in FIG. 12, a time differentiation section 802 of theanalysis unit 800 illustrated in FIG. 11 acquires a time differentiatedwaveform that is a waveform obtained by differentiating the basewaveform along the time axis. The time differentiated waveform isrepresented as the solid line on a graph as illustrated in FIG. 9 andFIG. 10. The relationship between the base waveform and the timedifferentiated waveform is similar to that of the first aspect asdescribed above.

Then, in a measurement value differentiation process S23 in the analysisprocess S20 illustrated in FIG. 12, a measurement value differentiationsection 803 in the analysis unit 800 illustrated in FIG. 11differentiates the base waveform along the optical measurement valueaxis, and acquires a measurement value differentiated waveform asillustrated by the dotted line in the graph of FIG. 10. The measurementvalue differentiated waveform is also similar to that of the firstaspect.

Then in a measurement value boundary determination process S24 in theanalysis process S20 illustrated in FIG. 12, a measurement valueboundary determination section 804 in the analysis unit 800 illustratedin FIG. 11 determines respective optical measurement values S1 to S6corresponding to the peaks T1′ to T6′ in FIG. 10 to be separationboundaries between components.

Then in an integral quantification process S28 in the analysis processS20 illustrated in FIG. 12, an integral quantification section 808 inthe analysis unit 800 illustrated in FIG. 11 determines the opticalmeasurement values S1 to S6, which respectively correspond to the peaksT1′ to T6′ determined in the measurement value boundary determinationprocess S24, to be separation boundaries, and then determines timepoints t1 to t6, which respectively correspond to the bottoms B1 to B6in the time differentiated waveform and respectively correspond to theseparation boundaries, to be integration boundaries. Then, for eachintegration interval whose both ends are adjacent two integrationboundaries, the time differentiated waveform is integrated to calculatean area occupied by the time differentiated waveform. The value obtainedas the area can be regarded as relative amount of the componentscorresponding to the integration intervals (namely, the componentappearing as a peak waveform in the time differentiated waveform).

An important point here is that, although the bottom B3 in FIG. 10 doesnot always appear as a distinct minimal value in the time differentiatedwaveform, the peak T3′ corresponding thereto in the measurement valuedifferentiated waveform can be distinctly identified as a so-calledshoulder peak in the graph. Therefore, the time t3 corresponding tobottom B3 can be distinctly determined to be the integration boundary t3corresponding to the separation boundary S3. Then, integrating the timedifferentiated waveform for an integration interval whose both ends arethe integration boundary t3 and the adjacent integration boundary t4enables the relative amount to be calculated for a peak waveform P3which does not always appear as a distinct peak. Of course, for theother peak waveforms P1, P2, P4 and P5 that do appear as distinct peaks,the respective relative amounts thereof can also be calculated byrespectively integrating the time differentiated waveform in a similarmanner using respective integration intervals of t1 to t2, t2 to t3, t4to t5, and t5 to t6.

Third Aspect

A component analysis method of a third aspect of the present disclosureis an augmented configuration of the component analysis method of thefirst aspect, wherein the analysis process S20 further includes a shiftquantification process S25 in which a distance of each interval whoseboth ends are adjacent two separation boundaries along the axis of theoptical measurement values is calculated as a relative amount of eachcomponent in the sample Sa corresponding to the interval.

The component analysis device 1 of the present aspect is an augmentedconfiguration of the component analysis device 1 of the first aspect,wherein the analysis unit 800 further includes a shift quantificationsection 805 that calculates a distance of each interval whose both endsare adjacent two separation boundaries along the axis of the opticalmeasurement values is calculated as a relative amount of each componentin the sample Sa corresponding to the respective intervals.

A functional configuration of the analysis unit 800 of the presentaspect is illustrated in FIG. 13. Explanation follows regarding thecomponent analysis method of the present aspect, with reference to aflowchart of FIG. 14.

The measurement process S10 illustrated in FIG. 14 is similar to that inthe first aspect.

In a base waveform acquisition process S21 in the analysis process S20illustrated in FIG. 14, a base waveform acquisition section 801 in theanalysis unit 800 illustrated in FIG. 13 acquires a base waveform byplotting the optical absorbance values as the optical measurement valuesalong one axis (the Y axis, for example) of a two-dimensional planeagainst a time axis as another axis (the X axis, for example). The basewaveform is, for example, represented as a solid line on a graph such asillustrated in FIG. 8. The base waveform acquisition process S21 issimilar to that of the first aspect.

Then, in a measurement value differentiation process S23 in the analysisprocess S20 illustrated in FIG. 14, a measurement value differentiationsection 803 in the analysis unit 800 illustrated in FIG. 13differentiates the base waveform along the optical measurement valueaxis to acquire a measurement value differentiated waveform such asillustrated by the dotted line on the graph in FIG. 15. The measurementvalue differentiated waveform is also similar to that of the firstaspect.

Next in a measurement value boundary determination process S24 in theanalysis process S20 illustrated in FIG. 14, a measurement valueboundary determination section 804 in the analysis unit 800 illustratedin FIG. 13 determines optical measurement values S1 to S6 respectivelycorresponding to the peaks T1′ to T6′ in FIG. 15 to be separationboundaries between components.

Then, in a shift quantification process S25 in the analysis process S20illustrated in FIG. 14, a shift quantification process 805 in theanalysis unit 800 illustrated in FIG. 13, without acquiring a timedifferentiated waveform, calculates relative amounts of components inthe sample Sa based on distances D1 to D5 that are intervals demarcatedby separation boundaries S1 to S6, which respectively correspond to thepeaks T1′ to T6′ determined in a similar manner to in the first aspect,as illustrated in FIG. 15. Note that the distances of intervals D1 to D5illustrated in FIG. 15 correspond to the areas of the peak waveforms P1to P5 illustrated in FIG. 10. In the shift quantification process S25,It is not necessary to refer to a time differentiated waveform at all.

Fourth Aspect

A component analysis method of a fourth aspect of the present disclosureincludes a measurement process S10 and an analysis process S20. In themeasurement process S10, a sample solution that is continuouslyintroduced into a flow path 27 is separated into a plurality ofcomponents inside the flow path 27 and is optically measured over timeat a measurement position 27A on the flow path 27 to obtain opticalmeasurement values. In the analysis process S20, the plurality ofcomponents contained in a sample Sa is analyzed based on the opticalmeasurement values. The analysis process S20 includes: a base waveformacquisition process S21 in which a base waveform is acquired by plottingthe optical measurement values along a time axis on a two-dimensionalplane; a time differentiation process S22 in which a time differentiatedwaveform is acquired, which is a waveform obtained by differentiatingthe base waveform along the time axis; a reciprocal differentiationprocess S26 in which a reciprocal differentiated waveform is acquired,which is a waveform obtained by plotting reciprocals of the timedifferentiated waveform along the time axis; and a time boundarydetermination process S27 in which time points corresponding to peaks inthe reciprocal differentiated waveform are determined to be separationboundaries between adjacent components in the plurality of components.

A component analysis device 1 of the present aspect includes: a flowpath 27 into which a sample solution is continuously introduced; ameasurement section 40 that optically measures the sample solution overtime, which is separated into a plurality of components inside the flowpath 27, at a measurement position 27A on the flow path 27 to obtainoptical measurement values; and an analysis unit 800 that analyzes theplurality of components contained in a sample Sa based on the opticalmeasurement values. The analysis unit 800 includes: a base waveformacquisition section 801 that acquires a base waveform by plotting theoptical measurement values along a time axis on a two-dimensional plane;a time differentiation section 802 that acquires a time differentiatedwaveform that is a waveform obtained by differentiating the basewaveform along the time axis; a reciprocal differentiation section 806that acquires a reciprocal differentiated waveform that is a waveformobtained by plotting reciprocals of the time differentiated waveformalong the time axis; and a time boundary determination section 807 thatdetermines time points corresponding to peaks in the reciprocaldifferentiated waveform to be separation boundaries between adjacentcomponents in the plurality of components.

A functional configuration of the analysis unit 800 of the presentaspect is illustrated in FIG. 16. Explanation follows regarding thecomponent analysis method of the present aspect, with reference to aflowchart of FIG. 17.

The measurement process S10 illustrated in FIG. 17 is similar to that inthe first aspect.

In a base waveform acquisition process S21 in the analysis process S20illustrated in FIG. 17, a base waveform acquisition section 801 in theanalysis unit 800 illustrated in FIG. 16 acquires a base waveform byplotting the optical absorbance values as the optical measurement valuesalong one axis (the Y axis, for example) of a two-dimensional planeagainst a time axis as another axis (the X axis, for example). The basewaveform is, for example, represented as a solid line on a graph such asillustrated in FIG. 8. The base waveform acquisition process S21 issimilar to that of the first aspect. Note that the base waveform may bereferred to as a function in which the optical measurement value isplotted with time as a variable.

Next, in a time differentiation process S22 in the analysis process S20illustrated in FIG. 17, a time differentiation section 802 of theanalysis unit 800 illustrated in FIG. 16 acquires a time differentiatedwaveform that is a waveform obtained by differentiating the basewaveform along the time axis. The time differentiated waveform isrepresented as the solid line on a graph as illustrated in FIG. 9. Therelationship between the base waveform and the time differentiatedwaveform is similar to that mentioned in the first aspect as describedabove.

Next, in a reciprocal differentiation process S26 in the analysisprocess S20 illustrated in FIG. 17, a reciprocal differentiation section806 of the analysis unit 800 illustrated in FIG. 16 acquires areciprocal differentiated waveform as illustrated by the dotted line inFIG. 18 by plotting the reciprocal of the optical absorbances of thetime differentiated waveform acquired in the time differentiationprocess S22 along the time axis. Note that, when plotting the reciprocalof the optical absorbance of the time differentiated waveform on atwo-dimensional plane having the optical measurement values as one axisand the time axis as the other axis, it is sufficient to plot relativevalues that are precise enough to clearly correspond to the timedifferentiated waveform, and it is not necessary to plot absolutevalues. Note that the reciprocals of the optical absorbances may also beacquired as data that are capable of being plotted on a two-dimensionalplane, and it is not necessary to actually draw a graph based on suchdata.

Note that the peaks and bottoms on the time differentiated waveformrespectively correspond to the bottoms and peaks on the reciprocaldifferentiated waveform, similarly to the measurement valuedifferentiated waveform described in the first aspect. Thus, asillustrated in FIG. 18, peaks T1″ to T6″ of the reciprocaldifferentiated waveform respectively correspond to bottoms B1 to B6 inthe time differentiated waveform. In other words, the bottoms B1 to B6in the time differentiated waveform appear distinctly as the respectivepeaks T1″ to T6″ in the reciprocal differentiated waveform.

Namely, in a time boundary determination process S27 in the analysisprocess S20 illustrated in FIG. 17, a time boundary determinationsection 807 in the analysis unit 800 illustrated in FIG. 16 determinesthe time points t1 to t6 respectively corresponding to the peaks T1″ toT6″ in the reciprocal differentiated waveform to be separationboundaries between components.

The configuration of the fourth aspect as described above enables thebottoms B1 to B6, which are hitherto needed to be discriminated as thetwo ends of a peak waveform in the time differentiated waveform, can berespectively identified as the respective peaks T1″ to T6″ of thereciprocal differentiated waveform. Then, it is possible for the timepoints t1 to t6 corresponding to the peaks T1″ to T6″ to be respectivelydetermined to be the separation boundaries between components containedin the sample Sa.

Fifth Aspect

A component analysis method of an fifth aspect of the present disclosureis an augmented configuration of the component analysis method of thefourth aspect, wherein the analysis process S20 further includes anintegral quantification process S28 in which a value obtained byintegrating the time differentiated waveform for an integration intervalwhose both ends are adjacent two integration boundaries, wherein timepoints corresponding to the separation boundaries are determined to beintegration boundaries for the time differentiated waveform, iscalculated as a relative amount of each component in the sample Sacorresponding to the integration intervals.

The component analysis device 1 of the present aspect is an augmentedconfiguration of the component analysis device 1 of the fourth aspect,wherein the analysis unit 800 further includes an integralquantification section 808 that calculates values obtained byintegrating the time differentiated waveform for an integration intervalwhose both ends are adjacent two integration boundaries, wherein timepoints corresponding to the separation boundaries are determined to beintegration boundaries for the time differentiated waveform, as arelative amount of the component in the sample Sa corresponding to theintegration intervals.

A functional configuration of the analysis unit 800 of the presentaspect is illustrated in FIG. 19. Explanation next follows regarding thecomponent analysis method of the present aspect, with reference to aflowchart of FIG. 20. However, the measurement process S10, and the basewaveform acquisition process S21, the time differentiation process S22,the reciprocal differentiation process S26, and the time boundarydetermination process S27 in the analysis process S20 illustrated inFIG. 20 are similar to those of the fourth aspect.

In the time boundary determination process S27 illustrated in FIG. 20,in FIG. 18, the time points t1 to t6 respectively corresponding to thepeaks T1″ to T6″ determined in the fourth aspect are the separationboundaries respectively corresponding to the bottoms B1 to B6, and thetime points t1 to t6 as these separation boundaries are respectivelydetermined to be integration boundaries. Then, in an integralquantification process S28 in the analysis process S20 illustrated inFIG. 20, an integral quantification section 808 in the analysis unit 800illustrated in FIG. 19 integrates the time differentiated waveform foreach integration interval whose both ends are adjacent two integrationboundaries, and thereby calculates an area occupied by the timedifferentiated waveform in each integration interval. The value obtainedfor the areas can be regarded as relative amount of componentscorresponding to the integration intervals (namely, the componentappearing as a peak waveform in the time differentiated waveform).

An important point here is that, although the bottom B3 in FIG. 18 doesnot always appear as a distinct minimal value in the time differentiatedwaveform, the peak T3″ corresponding thereto in the reciprocaldifferentiated waveform can be distinctly identified as a so-calledshoulder peak in the graph. Therefore, the time t3 corresponding tobottom B3 can be distinctly determined to be the integration boundaryt3. Then, integrating the time differentiated waveform for anintegration interval whose both ends are the integration boundary t3 andthe adjacent integration boundary t4 enables the relative amount to becalculated for a peak waveform P3 which does not always appear as adistinct peak. Of course, for the other peak waveforms Pb, P2, P4 and P5that do appear as distinct peaks, the respective relative amountsthereof can also be calculated by respectively integrating the timedifferentiated waveform in a similar manner using respective integrationintervals of t1 to t2, t2 to t3, t4 to t5, and t5 to t6.

Others

As another exemplary embodiment of the present invention, there are alsoexamples of component analysis methods other than capillaryelectrophoresis, such as one in which another method (for example,chromatography) is adopted to separate the sample solution when thesample solution is introduced into the flow path by using a methoddifferent to applying a voltage, and in which measurement is performedwith a flow path of larger width than the capillary tube describedabove.

EXAMPLES

An example of a case in which blood of an examinee is employed as thesample Sa will now be used to illustrate advantageous effects of thepresent exemplary embodiment with respect to the effects on identifyingpeak waveforms of high and low ambient temperature and high and lowconcentrations of a specimen during measurement.

FIG. 21A to FIG. 21D illustrate the effects of high and low ambienttemperature and high and low concentrations of a specimen duringmeasurement for a time differentiated waveform obtained by the timedifferentiation process S22. In FIG. 21A to FIG. 21D, each vertical axisrepresents differential values of absorbance. The ambient temperature inFIG. 21A and FIG. 21B was 23° C., and the ambient temperature in FIG.21C and FIG. 21D was 8° C. The sample Sa used in FIG. 21A and FIG. 21Cwas previously diluted by about three times (referred to as a lowconcentration sample), and the sample Sa used in FIG. 21B and FIG. 21Dis not diluted (referred to as a high concentration sample). In each ofthe drawings, a peak α corresponds to hemoglobin variant HbA1c, and aregion demarcated by distinct bottoms appearing at each end isillustrated by hatching.

At normal ambient temperature as in FIG. 21A and FIG. 21B, the bottomportion acting as the boundary to a peak β adjacent on the lowermigration speed side (namely, the right hand side) was distinct,irrespective of the high or low concentration of the sample Sa.

However, as illustrated in FIG. 21C and FIG. 21D, the peak α and thepeak β tended to approach each other when the ambient temperature was 8°C., making the boundary between the mutually adjacent peak α and peak βsomewhat indistinct. In particular, when there was a high concentrationsample as in FIG. 21D, the peak β was absorbed into the peak α andbecomes undiscernible. In cases in which an attempt is made to divide apeak waveform in such a state simply by using the distinct bottoms atthe two sides thereof, then a boundary to a peak γ even further to thelower migration speed side is used therefor, resulting in the regionapplied with hatching being used for quantification of the content ofthe peak α. However, this region actual includes the peak β. Thus, whena high concentration sample is measured at a low ambient temperature,the quantified content of the component corresponding to an identifiedpeak may possibly be calculated to be higher than is really the case.

FIG. 22A to FIG. 22D illustrate respective time differentiatedwaveforms, similarly to those of FIG. 21A to FIG. 21D. In FIG. 22A toFIG. 22D, each vertical axis represents differential values ofabsorbance. However, each of the peak α illustrated with hatching is aregion demarcated by integration boundaries determined by themeasurement value differentiation process S23 based on thenon-illustrated base waveform, the measurement value boundarydetermination process S24, and the integral quantification process S28.

The regions identified as the peak α in FIG. 22A, FIG. 22B, and FIG. 22Cdid not differ from the corresponding regions in FIG. 21A, FIG. 21B, andFIG. 21C. However, in FIG. 22D, a boundary between the peak α and thepeak β, which was fused with the peak as in FIG. 21D, could be madedistinct, enabling calculation of content quantification that appearedto be appropriate by comparing the identified region therein to the peakα in the other drawings.

It is thus apparent that the above exemplary embodiment enables acorrect peak waveform to be identified even in cases in which a highconcentration sample is measured at a low ambient temperature, such thatit is possible to correctly quantify the components contained in thesample.

INDUSTRIAL APPLICABILITY

The invention of the present application is applicable to a componentanalysis system using continuous sample introduction, and isparticularly applicable to a component analysis system using capillaryelectrophoresis.

What is claimed is:
 1. A component analysis method comprising: ameasurement process in which a sample solution that is continuouslyintroduced into a flow path is separated into a plurality of componentsinside the flow path and is optically measured over time at ameasurement position on the flow path to obtain optical measurementvalues; and an analysis process in which the plurality of componentscontained in a sample is analyzed based on the optical measurementvalues, the analysis process including: a base waveform acquisitionprocess in which a base waveform is acquired by plotting the opticalmeasurement values against a time axis on a two-dimensional plane; ameasurement value differentiation process in which a measurement valuedifferentiated waveform is acquired, which is a waveform obtained bydifferentiating the base waveform with respect to the opticalmeasurement value along an axis of the optical measurement values; and ameasurement value boundary determination process in which the opticalmeasurement values corresponding to peaks in the measurement valuedifferentiated waveform are determined to be separation boundariesbetween adjacent components of the plurality of components.
 2. Thecomponent analysis method of claim 1, wherein the analysis processfurther includes: a time differentiation process in which a timedifferentiated waveform is acquired, which is a waveform obtained bydifferentiating the base waveform along the time axis; and an integralquantification process in which, a value obtained by integrating thetime differentiated waveform for an integration interval whose both endsare adjacent two integration boundaries, wherein time pointscorresponding to the separation boundaries are determined to beintegration boundaries for the time differentiated waveform, iscalculated as a relative amount of each component in the samplecorresponding to the integration interval.
 3. The component analysismethod of claim 1, wherein the analysis process further includes a shiftquantification process in which a distance of each interval whose bothends are adjacent two separation boundaries along the axis of theoptical measurement values is calculated as a relative amount of eachcomponent in the sample corresponding to the interval.
 4. The componentanalysis method of claim 1, wherein the optical measurement values areobtained using capillary electrophoresis in which a voltage is appliedto the sample solution being continuously introduced into a capillarytube as the flow path so as to separate the plurality of components inthe sample solution.
 5. The component analysis method of claim 1,wherein the plurality of components includes hemoglobin.
 6. Thecomponent analysis method of claim 1, wherein the base waveform includesa high-gradient portion that indicates arrival of at least one of theplurality of components at the measurement position on the flow path. 7.A component analysis method comprising: a measurement process in which asample solution that is continuously introduced into a flow path isseparated into a plurality of components inside the flow path and isoptically measured over time at a measurement position on the flow pathto obtain optical measurement values; and an analysis process in whichthe plurality of components contained in a sample is analyzed based onthe optical measurement values, the analysis process including: a basewaveform acquisition process in which a base waveform is acquired byplotting the optical measurement values against a time axis on atwo-dimensional plane; a time differentiation process in which a timedifferentiated waveform is acquired, which is a waveform obtained bydifferentiating the base waveform with respect to time along the timeaxis; a reciprocal differentiation process in which a reciprocaldifferentiated waveform is acquired, which is a waveform obtained byplotting reciprocals of the time differentiated waveform along the timeaxis; and a time boundary determination process in which time pointscorresponding to peaks in the reciprocal differentiated waveform aredetermined to be separation boundaries between adjacent components inthe plurality of components.
 8. The component analysis method of claim7, wherein the analysis process further includes an integralquantification process in which a value obtained by integrating the timedifferentiated waveform for an integration interval whose both ends areadjacent two integration boundaries, wherein time points correspondingto the separation boundaries are determined to be integration boundariesfor the time differentiated waveform, is calculated as a relative amountof each component in the sample corresponding to the integrationintervals.
 9. The component analysis method of claim 7, wherein theplurality of components includes hemoglobin.
 10. The component analysismethod of claim 7, wherein the base waveform includes a high-gradientportion that indicates arrival of at least one of the plurality ofcomponents at the measurement position on the flow path.
 11. A componentanalysis device comprising: a flow path into which a sample solution iscontinuously introduced; and a processing unit comprising hardwareconfigured to execute a program to implement: a measurement section thatoptically measures the sample solution over time, which is separatedinto a plurality of components inside the flow path, at a measurementposition on the flow path to obtain optical measurement values; and ananalysis unit that analyzes the plurality of components contained in asample based on the optical measurement values, the analysis unitincluding: a base waveform acquisition section that acquires a basewaveform by plotting the optical measurement values against a time axison a two-dimensional plane; a measurement value differentiation sectionthat acquires a measurement value differentiated waveform that is awaveform obtained by differentiating the base waveform with respect tothe optical measurement value along an axis of the optical measurementvalues; and a measurement value boundary determination section thatdetermines optical measurement values corresponding to peaks in themeasurement value differentiated waveform to be separation boundariesbetween adjacent components into the plurality of components.
 12. Thecomponent analysis device of claim 11, wherein the analysis unit furtherincludes: a time differentiation section that acquires a timedifferentiated waveform that is a waveform obtained by differentiatingthe base waveform along the time axis; and an integral quantificationsection that calculates, a value obtained by integrating the timedifferentiated waveform for each integration interval whose both endsare adjacent two integration boundaries, wherein time pointscorresponding to the separation boundaries are determined to beintegration boundaries for the time differentiated waveform, as arelative amount of each component in the sample corresponding to theintegration interval.
 13. The component analysis device of claim 11,wherein the analysis unit further includes a shift quantificationsection that calculates a distance of each interval whose both ends areadjacent two separation boundaries along the axis of the opticalmeasurement values is calculated as a relative amount of each componentin the sample solution corresponding to the interval.
 14. The componentanalysis device of claim 11, wherein the plurality of componentsincludes hemoglobin.
 15. The component analysis device of claim 11,wherein the base waveform includes a high-gradient portion thatindicates arrival of at least one of the plurality of components at themeasurement position on the flow path.
 16. A component analysis devicecomprising: a flow path into which a sample solution is continuouslyintroduced; and a processing unit comprising hardware configured toexecute a program to implement: a measurement section that opticallymeasures the sample solution over time, which is separated into aplurality of components inside the flow path, at a measurement positionon the flow path to obtain optical measurement values; and an analysisunit that analyzes the plurality of components contained in a samplebased on the optical measurement values, the analysis unit including: abase waveform acquisition section that acquires a base waveform byplotting the optical measurement values against a time axis on atwo-dimensional plane; a time differentiation section that acquires atime differentiated waveform that is a waveform obtained bydifferentiating the base waveform with respect to time along the timeaxis; a reciprocal differentiation section that acquires a reciprocaldifferentiated waveform that is a waveform obtained by plottingreciprocals of the time differentiated waveform along the time axis; anda time boundary determination section that determines time pointscorresponding to peaks in the reciprocal differentiated waveform to beseparation boundaries between adjacent components in the plurality ofcomponents.
 17. The component analysis device of claim 16, wherein theanalysis unit further includes an integral quantification section thatcalculates a value obtained by integrating the time differentiatedwaveform for an integration interval whose both ends are adjacent twointegration boundaries, wherein time points corresponding to theseparation boundaries are determined to be integration boundaries forthe time differentiated waveform, as a relative amount of each componentin the sample corresponding to the integration interval.
 18. Thecomponent analysis device of claim 16, wherein the plurality ofcomponents includes hemoglobin.
 19. The component analysis device ofclaim 16, wherein the base waveform includes a high-gradient portionthat indicates arrival of at least one of the plurality of components atthe measurement position on the flow path.
 20. The component analysisdevice of claim 16, wherein the sample solution is separated into theplurality of components inside the flow path based on capillaryelectrophoresis.